1, 2, methylene androstane derivatives



United States Patent C) F ABSTRACT OF THE DISCLOSURE A steroid compoundhaving the general structural formula I Q2 Q3 wherein R is a memberselected from the group consisting of hydrogen and acyl, acyl beingderived from an aliphatic carboxylic acid having from 1 to 4 carbonatoms, and R is a member selected from the group consisting of loweralkyl, lower alkenyl and lower alkinyl containing from 1 to 4 carbonatoms.

An example of the steroids is1,2a-methylene-l7a-methyl-u-androstane-l7fl-ol.

The compound has a strong anabolic and ovulation in- Q hibiting action.

The invention relates to 1,2a-methylene-5u-androstane derivatives of thegeneral formula wherein R is a member selected from the group consistingof hydrogen and acyl, acyl being derived from an aliphatic, carboxylicacid having from 1 to 11 carbon atoms and R is a member selected fromthe group consisting of lower alkyl, lower alkenyl and lower alkinylcontaining from 1 to 4 carbon atoms.

It is an object of the present invention to improve the anabolicefliciency of steroids, particularly in case of oral 5 application.

Another object of the invention is to improve the ovulation inhibitoryactivity of androstane type steroids.

A more specific object of the invention is the provision of a methylenesubstituted androstane type steroid with marked anabolic and ovulationinhibiting action.

These and other objects, as will be apparent from the followingdescription of the invention, are accomplished by the compounds of thegeneral formula above stated. The acyl group that may be present as R inthis formula may consist of any of the acid residues that are commonlyused in steroid chemistry. Preferred acyl groups are the acid residuesof aliphatic carboxylic acids and particularly those having 1 to 11carbon atoms. Examples of such acids 'ice are acetic acid, propionicacid, caproic acid, enanthic acid and undecylic acid. These acidresidues can of course also be substituted in the usual manner. Asexamples in this respect are mentioned phenyl acetic acid,cyclopentylpropionic acid, halogenoacetic acid, amino-acetic acid,hydroxypropionic acid, etc.

7 The R groups in the above stated formula may particularly be thosehaving between 1 and 4 carbon atoms such as methyl, ethyl, ethinyl orvinyl.

The following Table 1 shows the strong oral anabolic action of the newcompounds in item IV, relating to the 1,20; methylene 17ccmethyl-Sa-andrOStane-l7fi-ol. The oral anabolic and androgenic activityof the orally administered compound was determined by comparison withthe corresponding activity of 17a-methyltestosterone (item I), and ofthe well known anabolic agents II and III. The tests were carried out bystandard methods using castrated rats and the Levator-ani and semiclevesicle test.

Table 1 clearly demonstrates the superiority of the new compounds inrespect of their anabolic efficiency. The eX- tremely high dissociationbetween anabolic and androgenic action is illustrated by the subsequentTable 2.

TABLE 2 Levator- Semicle D ose ani Vesicle N 0. Substance P.O Weight,Weight,

mg. rug/100 g. mg./100 g.

rat rat 17a-methyl-testosterone 12 x 44 370 l,2a-methylene-l7a 12 x 3 4359 methyl-5a-androstane-17fi-ol.

The 1,20: methylene-17ot-alkyl-5a-androstane-17,8-01 compounds of theinvention and their l7-esters, in addition to their anabolic action, arestrong ovulation inhibitors. If we compare for instance the specificcompound listed as IV with conventional ovulation inhibitors as shownbelow in Table 3 as items V to VIII we find a 10 to 3-0 times increasedactivity after oral application to normal female rats. The positive ornegative ovulation in these tests was determined by tube inspection.

The following table lists the specific does (W which results in omissionof ovulation in the case of of the test animals and peroraladministration.

TABLE 3 PO. mg./ animal/day No. Compound Wow (continued for four days) V17a-ethmyl-19-nor-testosterone 3 VI 17a-ethinyl-lQ-nor-testosterone- 3acetate. VII... 17a-ethinyl-A -estrene-l7fi-ol-3- 1-10 one. VIII6-0hlor-A -l7a-hydroxyprogesterone- 1-3 acetate. IV1,2a-rnethylene-17a-methyl-5a- 0.1

androstanc-UB-ol.

:3 The new 1,2a-methylene-5ot-androstane derivatives may be combined fortherapeutical use with the additives, vehicles, taste modifiers, etc.common in galenic pharmacy and may be prepared in the form of tablets,lozenges, capsules, pills or also as suspensions or solutions. Theselection of the concentration of active ingredient in drugs of thistype depends of course on the indication for which it is intended. Oralanabolic agents for instance should contain about 1 to 20 mg. andovulation inhibitors should contain about 0.5 to 5 mg. active substance.

Regarding the process of manufacture, the novel1,2otmethylene-5a-androstane derivatives may for instance be made in thefollowing manner. LZwmCthYlCIIE-Saandrostane-l7-one may be subjected toreaction with a Grignard agent in conventional manner in order tointroduce an alkyl or alkenyl group in the place of the 17 keto group.In a similar manner can an alkinyl group be introduced, for instance theethinyl group, which can then be reduced to the corresponding vinylgroup or to a saturated alkyl group. Depending on what type of R isdesired in the final compound there may additionally be an.esterification of the primarily formed steroid alcohols with thedesired acid or a reactive derivative of such acid. The term desiredacid should be understood as implying all .acids conventionally used insteroid chemistry for the purpose of esterification of steroid alcohols.

The starting product of this process the 1,2a-methylene-5a-androstane-17-one has not heretofore been described, .as far as weare aware. This compound can for instance be made as follows from1,Ze-methylene-Sot-androstanel7fi-ol-3-one-l7-acetate using conventionalmethods.

5 g. of 1,2wmethylene-5a-androstane-17,8-ol-3-one-17- ;acetate [Ben 93,1710 (60)] are heated for 20 minutes to 130 with 370 ml. ethylene-glycoland 25 ml. hydrazinhydrate of an 80% concentration. After cooling andaddition of a solution of 37 g. sodium hydroxide in 37 ml. water heatingis effected for two hours to 190 to 200 C. upon slow distillation. Thereaction mixture is then .again cooled and poured into a slightlyacidified sodium chloride solution. The precipitate that forms isfiltered off and dissolved in either. After washing until neutral and\drying the ether solution is concentrated by evaporation in vacuo. Theresidue is subjected to chromatography over silicagel (having 5% watercontent) with methylene chloride/ chloroform (in a ratio of 3 to l) andis triturated with pentane. There are obtained 2.74 g. of LZa-methyl-.ene-5a-androstane-17/3-01 having a melting point between 111.5 and 112C.

4.1 g. of this product are dissolved in 5 ml. acetone.

1.25 g. of chromium-(VI)-oxide in 4.7 ml. 8 N-sulfuric .acid are addedto the solution upon stirring and external cooling in ice water. Thesolution is then poured into an .aqueous sodium chloride solution and isextracted with methylene chloride. After neutral washing, drying andconcentration by evaporation of the methylene chloride phase there areleft 4.0 g. of 1,2u-methylene-5a-andro- .stane-17-one in the form of anoil.

The following examples will illustrate the making of the compounds ofthe invention without being intended .as any limitation of the scope ofthe invention.

Example 1 A Grignard solution was prepared from 4.49 g. magnesiumfilings in 5-0 ml. abs. ether and 11.6 ml. methyliodide in 32.4 ml. abs.ether. 4.0 g. LZOL-ITICthYlCIlB-Sotandrostane-17-one in 140 ml. abs.benzene were then added by dropping to the Grignard solution and werestirred for 4 hours at room temperature in a nitrogen atmosphere.Concentrated aqueous ammonium chloride solution was then cautiouslyadded to the reaction mixture upon cooling with ice. The product wasfinally slightly acidified with dilute hydrogen chloride and extractedwith ether. After separation the ether phase was washed until neutral,dried and concentrated by evaporation. The

residue was Subject d t chromatography over silicagel 4 (containing 5%water) with methylene chloride. There were obtained 3.06 g.1,2a-methylene-17u-rnethyl-5mandrostane-17/3-ol, M.P. 52-54 C. [111-4.85 (chloroform). NearlR:163 and 2.21; (cyclopropane); IR: 2.93, (OH),8.70/9.09/9.34,u. (COH).

Example 2 Acetylene was passed for 30 minutes upon cooling into 133 ml.abs. tetrahydrofurane, A Grignard solution was prepared from 9.26 g.magnesium filings in 114 ml. abs. tetrahydrofurane and 29 ml.ethylbromide in 114 ml. abs. tetrahydrofurane. The cooled Grignardsolution was added to the above solution of tetrahydrofurane resultingin a temperature rise to C. The introduction of acetylene was continueduntil the reaction temperature went down again. 4.8 g. of1,2a-methylene-5ot-and-rostane-17-one in 114 ml. abs. tetrahydrofuraneWere then drop-added while continuing the passing of acetylene throughthe mixture. The reaction product was thereafter heated and stirred for21 hours in an oil bath of a temperature of 70 C. and a nitrogenatmosphere. After cooling to 5 C. concentrated aqueous ammonium chloridesolution was slowly added until there was no more reaction.

The product was thereupon extracted with ether, the separated organicphase was washed neutral with water and after drying over sodium sulfatewas concentrated by evaporation in vacuo to dryness. The residue wassubjected to chromatography over silicagel. By elution with carbontetrachloride/methylenechloride (in a ratio of 2 to 1) one obtained 2.65g. of 1,Za-methylene-17a-ethinyl- Swandrostane-Ufi-Ol; M.P. 84-86 C.

[a.] 34.1 (chloroform). Near-JR: 1.63 and 2.21,!1. (cyclopropane); IR:2.95 (OH), 3.06 (CH of CECH), 4.77; (CEC), 9.55/9.65/9.86/9.93,u.(C-OH).

Example 3 The product was thereafter filtered ofi from the catalyst andthe solution was concentrated to dryness in vacuo. There were thusobtained 300 mg. 1,2a-methylene-17avinyl-5a-androstane-17B-ol in theform of an oil. [a] 2 +5.95 (chloroform) IR: 2.91 1. (OH), 6.1,u. (C C),9.82/10.0/10.90,u. (C-OH and CH=CH Example 4 312 mg. of1,2a-methylene-17a-ethinyl-5tx-androstanel7fi-ol were hydrogenated in 30ml. thiophene-free benzene in the presence of 2x312 mg. Lindlar catalystup to step-by-step absorption of 2 m. mole of hydrogen. The solution wasthen filtered off from the catalyst and was concentrated to dryness at40 in vacuo. There were thus obtained 300 mg. of1,2a-methylene-l7a-ethyl-5oc-androstane-l7B-ol in the form of an oil.[001 2 +0.95 (chloroform). IR: 2.87;. (OH), 9.87/10.27,u (C-OH).

Example 5 A mixture of 500 mg. Ila-methylene-l7m-ethinyl-5wandrostane-l7B-ol, 2.5 ml. pyridine and 2.5 ml. acetic acid anhydridewere heated for 4 hours to l40 C. The reaction mixture was then stirredinto an iced sodium chloride solution and was extracted with ether.After neutral washing, drying and concentration by evaporation of theseparated ether solution there were obtained 425 mg. 1,2oz methylene ae-thinyl 5a androstane 17B- ol-l7-acetate in the form of an oil.

All of these examples are intended for illustration only, as haspreviously been noted, and we therefore do not wish to be limitedotherwise than by the language of the appended claims.

5 We claim: 1. A compound having the general structural formula R, a I Iwherein R is a member selected from the group consisting of hydrogen andacyl, acyl being derived from an aliphatic carboxylic acid having from 1to 11 carbon atoms, 15

and R is a member selected from the group consisting of lower alkyl,lower alkenyl and lower alkinyl containing from 1 to 4 carbon atoms.

2. lint-methylene-17a-methyI-Sa andrOstane-17,8-01. 3.1,2a-methy1ene-17a-ethiny1-5a-androstane-17 3-01.

UNITED STATES PATENTS 3,163,663 12/1964 Cross et al 260-397.5 103,242,050 3/ 1966 Wiechert 260-397.5 3,254,099 5/1966 Jolles et a1260397.5

OTHER REFERENCES Fieser et a1.: Steroids, Reinhold Publishing Corp., NewYork, N.Y., 1959, pages 492, 493, 621, 627 and 630.

LEWIS GOTTS, Primary Examiner.

J. R. BROWN, Assistant Examiner. 20

1. A COMPOUND HAVING THE GENERAL STRUCTURAL FORMULA